CRISPR/Cas系统的概述及其在干扰基因表达研究中的应用及存在的问题  

The Overview of CRISPR/Cas System and Its Application and Existing Problems in the Research of Interfering Gene Expression

作　　者：赵秀玲[1] 黄愉淋[1] 潘宏[1] 张明[1] 

机构地区：[1]广西大学动物繁殖研究所,亚热带农业生物资源保护与利用国家重点实验室,南宁530004

出　　处：《基因组学与应用生物学》2016年第3期672-676,共5页Genomics and Applied Biology

基　　金：广西自然科学基金项目桂财教[2015]139号资助

摘　　要：CRISPR(clustered regularly interspaced short palindromic repeats)/Cas系统是源于原核生物的一种获得性免疫防御系统,与Cas蛋白协同作用保护细菌等免受病毒或者质粒的二次感染,广泛存在于细菌和古细菌中。科学家们通过改造CRISPR/Cas系统,使其成为一种新型的基因编辑工具。最近,科学家们又通过突变Cas9这个核酸酶的切割活性区域,使其成仍可结合到特定的核酸序列上,但却失去核酸切割活性的d Cas9(dead Cas9)蛋白,利用这个空间位阻效应达到抑制基因表达的效果,实现基因的沉默。这就是利用CRISPR系统干扰基因表达基本原理。本文主要综述CRISPR/Cas系统的结构、种类、作用机制及其在干扰基因表达中的应用和存在的问题。CRISPR（clustered regularly interspaced short palindromic repeats）/Cas system,which cooperate with Cas protein family to protect the bacteria from secondary infection of the virus or plasmid,is a kind of acquired immune defense system of prokaryote and widely found in bacteria and archaea. Scientists make it become a new kind of gene editing tool by reforming the CRISPR/Cas system. Recently,in order to inhibit the expression of genes and then carry out gene silencing using stereo-hindrance effect,scientists make it a d Cas9（dead Cas9） protein which can still combine to the specific nucleotide sequence but losing the cleavage activity of nuclease by mutating the cleavage activity region of Cas9. This is the basic principles of repressing expression of targeted genes by CRISPRi system. This review mainly summarized the structure,types,mechanism of CRISPR/Cas system,and its application and existing problems in interfering gene expression.

关 键 词：CRISPR/Cas9 干扰 dCas9 基因表达 

分 类 号：Q78[生物学—分子生物学]